四翅滨藜BADH基因的克隆、序列分析及其原核表达  

Cloning and Analysis of A BADH Gene from Atriplex canescens and Its Prokaryotic Expression

作　　者：张勇[1] 余刚[1] 冯俊[1] 毕玉[1] 迪娜.塔布斯 潘洪玉[1] 

机构地区：[1]吉林大学植物科学学院,长春130062

出　　处：《吉林农业大学学报》2012年第6期618-623,共6页Journal of Jilin Agricultural University

基　　金：农业部转基因生物新品种培育重大专项(2009ZX08009-062B)

摘　　要：在构建四翅滨藜(Atriplex canescens)全长cDNA文库中通过随机克隆测序并进行EST分析基础上,得到1个四翅滨藜甜菜碱醛脱氢酶(betaine aldehyde dehydrogenase,BADH)的cDNA序列,命名为AcBADH。AcBADH cDNA包含1个长度为1 500 bp的完整开放阅读框,编码500个氨基酸,属于ALDH-SF超家族,其核酸序列与中亚滨藜的BADH基因的同源性为98%,对应编码氨基酸序列同源性为99%。将得到的序列提交GenBank(登录号:JF776157)。与其他植物BADH的氨基酸序列比对,AcBADH具有相同功能区域,包括N-端信号肽,底物结合及酶催化位点,因而推测可以参与甜菜碱的合成反应。对AcBADH与其他植物BADH的氨基酸序列的进化分析表明,AcBADH与鞑靼滨藜、中亚滨藜的亲缘关系较近。将AcBADH基因与原核表达载体pET28a连接,进行融合表达,在大肠杆菌BL21(DE3)中诱导表达出分子质量约58 kD的蛋白。A betaine aldehyde dehydrogenase gene was isolated from the library of Atriplex canescens,and its EST was analyzed.It was named as AcBADH.The full length of AcBADH contained an open reading frame of 1 500 bp.It encoded a polypeptide of 500 amino acids,belonging to ALDH-SF superfamily.The AcBADH shared 98% and 99% homology in nucleotide and amino acid to BADH from Atriplex centralasiatica,respectively.The accession number of AcBADH in GenBank was JF776157.Comparison of amino acid sequence with BADH from various plants showed that AcBADH possessed the same functional regions.Phylogenic analysis on the amino acid sequence of AcBADH with other plants showed that Atriplex canescens was closely related to Atriplex tatarica and Atriplex centralasiatica.AcBADH was ligated into the prokaryotic expression vector of pET28a and expressed with fusion protein（about 58 kD） in Escherichia coli BL2l（DE3）.

关 键 词：四翅滨藜 甜菜碱醛脱氢酶 基因克隆 序列分析 原核表达 

分 类 号：Q785[生物学—分子生物学] Q786